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Background: Cholangiocarcinoma (CCA) is a variety of biliary epithelial tumors involving intrahepatic, perihilar and distal bile duct. It is the most common malignant bile duct tumor in the liver and the second most common primary liver cancer, whose molecular mechanism not fully understood. Speci cally, the relationship between CCA and chondroitin polymerizing factor (CHPF) is still not clear. Methods: In this study, detection of clinical specimens was performed to preliminarily study the role of CHPF in cholangiocarcinoma. Cholangiocarcinoma cells with CHPF knockdown were constructed for in vitro study, which was also used in the construction of mice xenograft model for investigating the role of CHPF in the development of cholangiocarcinoma. Results: The results demonstrated that CHPF was signi cantly upregulated in cholangiocarcinoma tissues compared with normal tissues. High expression of CHPF was correlated with more advanced tumor grade. Moreover, knockdown of CHPF signi cantly inhibited cell proliferation, cell migration, promoted cell apoptosis and arrest cell cycle in G2 phase in vitro, as well as suppressed tumor growth in vivo. Conclusions: In conclusion, CHPF was identi ed as a tumor promotor in the development and metastasis of cholangiocarcinoma, which may provide a novel therapeutic target for the targeted therapy against cholangiocarcinoma.